GCLM (NM_002061) Human Tagged ORF Clone

CAT#: RC207224

GCLM (Myc-DDK-tagged)-Human glutamate-cysteine ligase, modifier subunit (GCLM)

Product Images

HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY GCLM (Cat# RC207224, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-GCLM(Cat# TA590111). Positive lysates LY400752 (100ug) and LC400752 (20ug) can be purchased separately from OriGene.

• 

  Western blot validation of overexpression lysate (Cat# LY400752) using anti-DDK antibody (Cat# TA50011-100). Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC207224 using transfection reagent MegaTran 2.0 (Cat# TT210002).

• 

  Coomassie blue staining of purified GCLM protein (Cat# TP307224). The protein was produced from HEK293T cells transfected with GCLM cDNA clone (Cat# RC207224) using MegaTran 2.0 (Cat# TT210002).

Specifications

Product Data

• Type: Human Tagged ORF Clone
• Tag: Myc-DDK
• Symbol: GCLM
• Synonyms: GLCLR
• Vector: pCMV6-Entry
• E. coli Selection: Kanamycin (25 ug/mL)
• Mammalian Cell Selection: Neomycin
• Sequence Data:
  ORF Nucleotide Sequence

  >RC207224 ORF sequence
  Red=Cloning site Blue=ORF Green=Tags(s)
  
  TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC
  GCCGCGATCGCC
  
  ATGGGCACCGACAGCCGCGCGGCCAAGGCGCTCCTGGCGCGGGCCCGCACCCTGCACCTGCAGACGGGGA
  ACCTGCTGAACTGGGGCCGCCTGCGGAAGAAGTGCCCGTCCACGCACAGCGAGGAGCTTCATGATTGTAT
  CCAAAAAACCTTGAATGAATGGAGTTCCCAAATCAACCCAGATTTGGTCAGGGAGTTTCCAGATGTCTTG
  GAATGCACTGTATCTCATGCAGTAGAAAAGATAAATCCTGATGAAAGAGAAGAAATGAAAGTTTCTGCAA
  AACTGTTCATTGTAGAATCAAACTCTTCATCATCAACTAGAAGTGCAGTTGACATGGCCTGTTCAGTCCT
  TGGAGTTGCACAGCTGGATTCTGTGATCATTGCTTCACCTCCTATTGAAGATGGAGTTAATCTTTCCTTG
  GAGCATTTACAGCCTTACTGGGAGGAATTAGAAAACTTAGTTCAGAGCAAAAAGATTGTTGCCATAGGTA
  CCTCTGATCTAGACAAAACACAGTTGGAACAGCTGTATCAGTGGGCACAGGTAAAACCAAATAGTAACCA
  AGTTAATCTTGCCTCCTGCTGTGTGATGCCACCAGATTTGACTGCATTTGCTAAACAATTTGACATACAG
  CTGTTGACTCACAATGATCCAAAAGAACTGCTTTCTGAAGCAAGTTTCCAAGAAGCTCTTCAGGAAAGCA
  TTCCTGACATTCAAGCGCACGAGTGGGTGCCGCTGTGGCTACTGCGGTATTCGGTCATTGTGAAAAGTAG
  AGGAATTATCAAATCAAAAGGCTACATTTTACAAGCTAAAAGAAGGGGTTCT
  
  ACGCGTACGCGGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATT
  ACAAGGATGACGACGATAAGGTTTAA
  Protein Sequence

  >RC207224 protein sequence
  Red=Cloning site Green=Tags(s)
  
  MGTDSRAAKALLARARTLHLQTGNLLNWGRLRKKCPSTHSEELHDCIQKTLNEWSSQINPDLVREFPDVL
  ECTVSHAVEKINPDEREEMKVSAKLFIVESNSSSSTRSAVDMACSVLGVAQLDSVIIASPPIEDGVNLSL
  EHLQPYWEELENLVQSKKIVAIGTSDLDKTQLEQLYQWAQVKPNSNQVNLASCCVMPPDLTAFAKQFDIQ
  LLTHNDPKELLSEASFQEALQESIPDIQAHEWVPLWLLRYSVIVKSRGIIKSKGYILQAKRRGS
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Restriction Sites: SgfI-MluI Cloning Scheme for this gene Plasmid Map
• ACCN: NM_002061
• ORF Size: 822 bp
• OTI Disclaimer: Due to the inherent nature of this plasmid, standard methods to replicate additional amounts of DNA in E. coli are highly likely to result in mutations and/or rearrangements. Therefore, OriGene does not guarantee the capability to replicate this plasmid DNA. Additional amounts of DNA can be purchased from OriGene with batch-specific, full-sequence verification at a reduced cost. Please contact our customer care team at custsupport@origene.com or by calling 301.340.3188 option 3 for pricing and delivery.
  
  The molecular sequence of this clone aligns with the gene accession number as a point of reference only. However, individual transcript sequences of the same gene can differ through naturally occurring variations (e.g. polymorphisms), each with its own valid existence. This clone is substantially in agreement with the reference, but a complete review of all prevailing variants is recommended prior to use. More info
• OTI Annotation: This clone was engineered to express the complete ORF with an expression tag. Expression varies depending on the nature of the gene.
• Product Components: The ORF clone is ion-exchange column purified and shipped in a 2D barcoded Matrix tube containing 10ug of transfection-ready, dried plasmid DNA (reconstitute with 100 ul of water).
• Reconstitution: 1. Centrifuge at 5,000xg for 5min.
  2. Carefully open the tube and add 100ul of sterile water to dissolve the DNA.
  3. Close the tube and incubate for 10 minutes at room temperature.
  4. Briefly vortex the tube and then do a quick spin (less than 5000xg) to concentrate the liquid at the bottom.
  5. Store the suspended plasmid at -20°C. The DNA is stable for at least one year from date of shipping when stored at -20°C.

Reference Data

• RefSeq: NM_002061.4
• RefSeq Size: 3074 bp
• RefSeq ORF: 825 bp
• Locus ID: 2730
• UniProt ID: P48507
• Cytogenetics: 1p22.1
• Protein Families: Druggable Genome
• Protein Pathways: Glutathione metabolism, Metabolic pathways
• MW: 30.7 kDa
• Gene Summary: Glutamate-cysteine ligase, also known as gamma-glutamylcysteine synthetase, is the first rate limiting enzyme of glutathione synthesis. The enzyme consists of two subunits, a heavy catalytic subunit and a light regulatory subunit. Gamma glutamylcysteine synthetase deficiency has been implicated in some forms of hemolytic anemia. Alternative splicing results in multiple transcript variants encoding different isoforms. [provided by RefSeq, Apr 2015]